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Description: Cellulose nanocrystals (CNCs) are gaining interest as a “green” nanomaterial with superior mechanical and chemical properties for high-performance nanocomposite materials; however, there is a lack of accurate material property characterization of individual CNCs. Here, a detailed study of the topography, elastic and adhesive properties of individual wood-derived CNCs is performed using atomic force microscopy (AFM). AFM experiments involving high-resolution dynamic mode imaging and jump-mode measurements were performed on individual CNCs under ambient conditions with 30% relative humidity (RH) and under a N2 atmosphere with 0.1% RH. A procedure was also developed to calculate the CNC transverse elastic modulus (EΤ) by comparing the experimental force-distance curves measured on the CNCs with 3D finite element calculations of tip indentation on the CNC. The EΤ of an isolated CNC was estimated to be between 18 and 50 GPa at 0.1% RH; however, the associated crystallographic orientation of the CNC could not be determined. CNC properties were reasonably uniform along the entire CNC length, despite variations along the axis of 3-8 nm in CNC height. The range of RH used in this study was found to have a minimal effect on the CNC geometry, confirming the resistance of the cellulose crystals to water penetration. CNC flexibility was also investigated by using the AFM tip as a nanomanipulator.